Anti-EEA1抗体- Early Endosome Marker
参阅全部 EEA1 一抗
兔多克隆抗体to EEA1 - Early Endosome Marker
Rabbit
Detects a band at 180kDa that represents EEA1 in Western blotting on human cell lines (corresponds to results seen in Mu et al). Also detects a band at 100kDa, we are unsure as to the identity of this band. Immunofluorescence staining of EEA1 in HeLa cells yields a punctate staining pattern consistent with the cytoplasmic distribution of endosomes. From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help. You may also be interested in our alternative recombinant antibody, ab109110.
适用于: WBmore details
不适用于: ICC/IF
与反应: Mouse, Rat, Dog, Human, Xenopus laevis, Chinese hamster
预测可用于: Chicken, Hamster, Cow, Zebrafish, Rhesus monkey, Aplysia
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab2900于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (12) | Use a concentration of 1 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 160 kDa). Abcam recommends using milk as the blocking agent. |
应用说明
Is unsuitable for ICC/IF.
Entrez Gene: 8411 Human
Entrez Gene: 216238 Mouse
Omim: 605070 Human
SwissProt: Q15075 Human
SwissProt: Q8BL66 Mouse
Unigene: 567367 Human
Unigene: 210035 Mouse
EEA 1 antibody
EEA1 antibody
EEA1_HUMAN antibody
Endosome associated protein p162 antibody
Endosome-associated protein p162 antibody
MST105 antibody
MSTP105 antibody
ZFYVE2 antibody
Zinc finger FYVE domain containing protein 2 antibody
Zinc finger FYVE domain-containing protein 2 antibody
Early endosome antigen 1 antibody
Early endosome antigen 1, 162kD antibody
Early endosome associated protein antibody
Western blot - Anti-EEA1 antibody - Early Endosome Marker (ab2900)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: EEA1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: NIH3T3 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab2900 observed at 162 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab2900 was shown to recognize EEA1 in wild-type HAP1 cells as signal was lost at the expected MW in EEA1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and EEA1 knockout samples were subjected to SDS-PAGE. Ab2900 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-EEA1 antibody - Early Endosome Marker (ab2900)
All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lane 4 : HeLa whole cell lysate
Lane 5 : HEK-293 whole cell lysate
Lane 6 : NIH 3T3 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG H&L (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 180 kDawhy is the actual band size different from the predicted?
Additional bands at: 100 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds
This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin (Lanes 1-3) or 3% Milk (Lanes 4-6) before being incubated with ab2900 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
Western blot - Anti-EEA1 antibody - Early Endosome Marker (ab2900)
All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa Fluor anti rabbit at 1/50000 dilution
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 180 kDawhy is the actual band size different from the predicted?
Additional bands at: 100 kDa, 41 kDa, 50 kDa. We are unsure as to the identity of these extra bands.
Fluorescence detection of secondary antibody.
Western blot - Anti-EEA1 antibody - Early Endosome Marker (ab2900)
All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1/500 dilution
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HEK293 Whole Cell lysate with Human EEA1 peptide (ab14946) at 1 µg
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 180 kDawhy is the actual band size different from the predicted?
Lane 1 - 2 : EEA1 antibody - Early Endosome Marker (ab2900) at 1/500 dilution, HEK293 Whole Cell lysate at 20 ug Lane 1 : as above Lane 2 : EEA1 peptide (ab14946) at 1 ug Secondary Goat polyclonal to Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution Performed under reducing conditions. Predicted band size : 160kD Observed band size : 180kD (why is the actual band size different from the predicted?)
Western blot - Anti-EEA1 antibody - Early Endosome Marker (ab2900)
All lanes : Anti-EEA1 antibody - Early Endosome Marker (ab2900) at 1 µg/ml
Lane 1 : Xenopus laevis lysate
Lane 2 : Mouse 3T3 cell lysate
Lane 3 : Mouse brain cell lysate
Lane 4 : Mouse liver cell lysate
Lane 5 : Rat brain cell lysate
Lane 6 : Rat liver cell lysate
Lane 7 : Dog lysate
Lane 8 : CHO cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 100,180 kDawhy is the actual band size different from the predicted?
Exposure time: 30 seconds
The Western blot shows that ab14944 reacts strongly with mouse 3T3 and CHO cell lysates. Weak cross-reactivity is seen with Xenopus, mouse brain, mouse liver, rat brain and dog lysates. The antibody does not appear to cross-react with rat liver lysate.
